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      LncRNA ADAMTS9‐AS2 suppresses the proliferation of gastric cancer cells and the tumorigenicity of cancer stem cells through regulating SPOP

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          Abstract

          Nowadays, research on CSCs is still in an initial stage, and there are few studies reporting the successful isolation and identification of CSCs. In the present study, we attempted to isolate CSCs through cultivating the cell line MKN45 in defined serum‐free medium and study the expression of stem cell markers or related proteins (Oct3/4, Sox2, Nanog and CD44) in CSCs. Moreover, immunofluorescence staining was performed to validate the stem cell markers of spheroid body‐forming cells. Further experiments were used to evaluate the SPOP expression in tumorsphere cells. In addition, ADAMTS9‐AS2 is a lncRNA that contributes to the genesis and development of many cancers, including gastric cancer (GC). We found ADAMTS9‐AS2 functioned as an anti‐oncogene and positively correlated with the expression of SPOP in GC tissues by combining bioinformatics analyses. Furthermore, we reported that ADAMTS9‐AS2 regulated the expression of SPOP in GC cells and tumorsphere cells to inhibit GC progression. Together, our results demonstrated that SPOP and ADAMTS9‐AS2 can be potential targets for GC treatment.

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          Most cited references21

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          RNA maps reveal new RNA classes and a possible function for pervasive transcription.

          Significant fractions of eukaryotic genomes give rise to RNA, much of which is unannotated and has reduced protein-coding potential. The genomic origins and the associations of human nuclear and cytosolic polyadenylated RNAs longer than 200 nucleotides (nt) and whole-cell RNAs less than 200 nt were investigated in this genome-wide study. Subcellular addresses for nucleotides present in detected RNAs were assigned, and their potential processing into short RNAs was investigated. Taken together, these observations suggest a novel role for some unannotated RNAs as primary transcripts for the production of short RNAs. Three potentially functional classes of RNAs have been identified, two of which are syntenically conserved and correlate with the expression state of protein-coding genes. These data support a highly interleaved organization of the human transcriptome.
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            Structures of SPOP-substrate complexes: insights into molecular architectures of BTB-Cul3 ubiquitin ligases.

            In the largest E3 ligase subfamily, Cul3 binds a BTB domain, and an associated protein-interaction domain such as MATH recruits substrates for ubiquitination. Here, we present biochemical and structural analyses of the MATH-BTB protein, SPOP. We define a SPOP-binding consensus (SBC) and determine structures revealing recognition of SBCs from the phosphatase Puc, the transcriptional regulator Ci, and the chromatin component MacroH2A. We identify a dimeric SPOP-Cul3 assembly involving a conserved helical structure C-terminal of BTB domains, which we call "3-box" due to its facilitating Cul3 binding and its resemblance to F-/SOCS-boxes in other cullin-based E3s. Structural flexibility between the substrate-binding MATH and Cul3-binding BTB/3-box domains potentially allows a SPOP dimer to engage multiple SBCs found within a single substrate, such as Puc. These studies provide a molecular understanding of how MATH-BTB proteins recruit substrates to Cul3 and how their dimerization and conformational variability may facilitate avid interactions with diverse substrates.
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              Hierarchical organization of prostate cancer cells in xenograft tumors: the CD44+alpha2beta1+ cell population is enriched in tumor-initiating cells.

              Prostate cancer cells are heterogeneous in their tumorigenicity. For example, the side population cells isolated from LAPC9 xenografts are 100 to 1,000 times more tumorigenic than the corresponding non-side population cells. Highly purified CD44(+) prostate cancer cells from several xenografts are also enriched in prostate cancer stem/progenitor cells. Because the CD44(+) prostate cancer cell population is still heterogeneous, we wonder whether we could further enrich for tumorigenic prostate cancer cells in this population using other markers. Integrin alpha2beta1 has been proposed to mark a population of normal human prostate stem cells. Therefore, we first asked whether the alpha2beta1(+/hi) cells in prostate tumors might also represent prostate cancer stem cells. Highly purified (> or =98%) alpha2beta1(+/hi) cells from three human xenograft tumors, Du145, LAPC4, and LAPC9, show higher clonal and clonogenic potential than the alpha2beta1(-/lo) cells in vitro. However, when injected into the nonobese diabetic/severe combined immunodeficient (NOD/SCID) mouse prostate or s.c., the alpha2beta1(+/hi) prostate cancer cells are no more tumorigenic than the alpha2beta1(-/lo) cells. Immunofluorescence studies reveal that CD44 and alpha2beta1 identify an overlapping and inclusive population of prostate cancer cells in that approximately 70% of alpha2beta1(+/hi) cells are CD44(+) and 20% to 30% of CD44(+) cells are distributed in the alpha2beta1(-/lo) cell population. Subsequently, we sorted out CD44(+)alpha2beta1(+/hi), CD44(+)alpha2beta1(-/lo), CD44(-)alpha2beta1(+/hi), and CD44(-)alpha2beta1(-/lo) cells from LAPC9 tumors and carried out tumorigenicity experiments. The results revealed a hierarchy in tumorigenic potential in the order of CD44(+)alpha2beta1(+/hi) approximately CD44(+)alpha2beta1(-/lo) > CD44(-)alpha2beta1(+/hi) > CD44(-)alpha2beta1(-/lo). These observations together suggest that prostate cancer cells are organized as a hierarchy.
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                Author and article information

                Contributors
                gaoxuesong@21cn.com
                xujunfei@ntu.edu.cn
                Journal
                J Cell Mol Med
                J. Cell. Mol. Med
                10.1111/(ISSN)1582-4934
                JCMM
                Journal of Cellular and Molecular Medicine
                John Wiley and Sons Inc. (Hoboken )
                1582-1838
                1582-4934
                11 March 2020
                April 2020
                : 24
                : 8 ( doiID: 10.1111/jcmm.v24.8 )
                : 4830-4838
                Affiliations
                [ 1 ] Department of General Surgery Affiliated Hospital of Nantong University Nantong China
                [ 2 ] Department of General Surgery Affiliated Hospital 2 of Nantong University Nantong China
                [ 3 ] Department of General Surgery Huai’an First People's Hospital Huai'an Jiangsu China
                [ 4 ] Medical College of Nantong University Nantong China
                Author notes
                [*] [* ] Correspondence

                Junfei Xu, Department of General Surgery, Affiliated Hospital of Nantong University, No. 20, Xisi Road, Nantong 226001, Jiangsu, China.

                Email: xujunfei@ 123456ntu.edu.cn

                Xuesong Gao, Department of General Surgery, Affiliated Hospital 2 of Nantong University, No. 6, Haier Lane North Road, Nantong 226001, Jiangsu, China.

                Email: gaoxuesong@ 12345621cn.com

                Author information
                https://orcid.org/0000-0002-1790-7130
                Article
                JCMM15161
                10.1111/jcmm.15161
                7176847
                32160650
                61d76621-1e18-4537-bba1-60a79a306d5c
                © 2020 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 19 August 2019
                : 11 February 2020
                : 25 February 2020
                Page count
                Figures: 5, Tables: 0, Pages: 9, Words: 5432
                Funding
                Funded by: Nantong Science and Technology Bureau , open-funder-registry 10.13039/501100009156;
                Award ID: MS12017001‐8
                Award ID: MS12018060
                Funded by: Clinical Basic Research Project of Nantong University
                Award ID: 2019JY004
                Categories
                Original Article
                Original Articles
                Custom metadata
                2.0
                April 2020
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.8.1 mode:remove_FC converted:23.04.2020

                Molecular medicine
                adamts9‐as2,cancer stem cells,gastric cancer,speckle‐type poz protein,tumorsphere

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